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Good afternoon, I am trying to make 18 % Acrylamide gel following...

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Shammi Akter Jul 2, 2020
protocol Protocol: Very Low Molecular Weight Proteins Electrophoresis Protocol
Good afternoon, I am trying to make 18 % Acrylamide gel following your protocol. Problem is 30 µl Teamed for 10 ml separating gel is quite high, because of it immediately going to polymerized in the tube. So, can I decreased teamed amount? My another question is, why you used 5 M (3 g) ur Read more Read less more less
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José David Rosales Author Answered Jul 7, 2020

Antibodies Massive Inmunoproduction Center, IDEA, Venezuela, Venezuela,

Good morning, you can use less Temed and thus decrease the polymerization time of acrylamity. You can test in a separate tube and time the amount of Temed and polymerization time, before using the crystals. The high concentration of urea helps to define the protein band. Otherwise the band is very diffuse. In effect it can load more volume of your protein and thus increase the possibilities of viewing it. Luck!!!
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